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VOL. 4, NO. 2, pp. 119-122 , May, 2014

Isolation and Identification Of Triterpenoid Saponin

From Baringtonia asiatica Kurz Seeds
Meity N Tanor1, Abdul Latief Abadi2, Bambang Tri Rahardjo2, Jantje Pelealu3
Doctoral Program, Faculty of Agriculture, University of Brawijaya, Indonesia
Lecturer of Plant Pest and Disease Major, Faculty of Agriculture, University of Brawijaya, Indonesia
Lecturer of Plant Pest and Disease Major, Sam Ratulangi University, Indonesia


The study aimed to identify the content of the triterpenoid saponin compound from Barringtonia
asiatica seeds collected from coastal of Malalayang beach, Manado, North Sulawesi. The method of
extraction was the maceration with methanol solvent. The identification of the isolate was by thin layer
chromatography, column chromatography, and GCMS. The phytochemistry test result of B. asiatica seed
extract contained the compounds of alkaloid, saponin, and tannin. Then, the compound was separated
by a thin layer chromatography method with a solvent system of methanol:chloroform:water. It
produced three nodes that were spread around polar (Rf 0.24), semipolar (Rf 0.6) and non -polar (Rf
0.78) areas. Meanwhile, the process of column chromatography could only separate two chemical com -
ponents namely semipolar (Rf 0.6) and polar (Rf 0.76). The identification with GCMS resulted in three
compounds of Triterpenoid saponins, namely 2.4-bis-(1.1-dimethyl ethyl)-, methylcarbamate; 4-
Dodecylphenol; and 2.6 bis-(1.1-dimethylethyl)-4-methyl-, methylcarbamate.

Keywords: isolation, identification, triterpenoid, Baringtonia asiatica.

INTRODUCTION is not toxic. Therefore, the study to identify the

content and the type of saponin resulted from B.
Barringtonia asiatica is a kind of mangrove plant asiatica seeds are significant. This information is
called “Sea Poison Tree” or “Fish Killer Tree." required for the supply of raw material in produ-
As its label says, this type of mangrove can insti- cing economical bio-pesticide. Since B. asiatica
gate toxicity even death of fish which consume grow in sandy and rocky aquaculture [12], this
its seeds [5]. However, B. asiatica has many bene- condition is mostly found in Indonesia.
fits. For example, its leaves are able to treat sto-
mach ache and be anti-rheumatic medication. Al- MATERIALS AND METHODS
so, its seeds are able to get rid of worms in the
intestine [12], be antifungal [6], be anti-inflamma- Formulating the crude extract of alkalo-
tion and analgesic [11], be anti-oxidant and anti- id, Saponin, Tannin dan Terpenoid tests
plasmodial [9]. Thus, the plant is expected to Samples of B. asiatica seeds was cleanly wa-
contain a saponin compound.
Saponin is included as a natural compound shed with running water and mashed with a blen-
with large molecule mass and value [7, 8, 1, 2]. der. 450 grams of B. asiatica seed powder was ma-
[10] state that saponin taken from Chenopodium cerated in a closed vessel for 3x24 hours and was
quinoa seeds is pesticide and its work mechanism- occasionally shaken in a shaker containing metha-
nol solvent. The solvent was filtrated by filter pa-
*Corresponding author: per and was evaporated afterwards in a rotary
Meity N Tanor
Doctoral Program, Faculty of Agriculture, University of
evaporator within the temperature of 35-37oC
Brawijaya, Indonesia (48-50 rpm). After that, we analyzed the content
E-mail: of alkaloid, flavanoid, and saponin based on [4].

JTLS | J. Trop. Life. Science 119 Volume 4 | Number 2 | May | 2014

Isolation and Identification Of Triterpenoid Saponin

Alkaloid Test was performed by adding 0.1 ture was 305,000C; the carrier gas, helium (He),
gram of the extract into 3 mL of chloroform and the speed of gas flow were 25,9 cm/sec; the pres-
3 drops of ammoniac. The chloroform fraction sure of carrier gas was 13,7 kPa; the column type
was separated and acidified by ten drops of H2- was Rastek Rxi-5MS; the column temperature
SO4 2M. The acid fraction, then, was added with was 70,00C; the ionization system was Electron
impact (EI), and the ionization energy was 70 Ev.
Meyer and Wagner reagents. The presence of al-
kaloid was evident when the white deposit was
made by Meyer reagent and the brown deposit by RESULTS AND DISCUSSION
Wegner reagent.
Saponin Test was performed by putting 1 The result of B. asiatica seed extract produced
gram of extract into a beaker containing 100 mL brownish white, oily, semi-solid compound in the
of water, and was boiled for 5 minutes. Then, it pasta form within the room temperature. The
was filtered, and the filtrate was tested. Ten mL result of phytochemistry test by [4] method con-
of filtrate was incubated in a closed test tube for firmed that the extract of B. asiatica seeds con-
10 minutes. The occurrence of saponin was iden- tained compounds of alkaloid, saponin, triterpe-
tified when stable froth/foam formation was ma- noid and tannin. From the test of Thin Layer
de. Crhomatography, there were three nodes in the
Tannin test was performed by mixing 0.1 polar (Rf score 0.24), semipolar (Rf score 0.6),
gram of extract with 2 mL of water and boiled and nonpolar (Rf score 0.78) areas. On the other
for 3 minutes. It was filtered, and the filtrate was hand, the method of Column Chromatography
added with a drop of FeCl3 1 % (b/v). The dark could only separate two chemistry components
blue or black colour exposed the existence of concentrated in the semipolar (Rf score 0.6) and
tannin. polar (Rf score 0.76) areas (Table 1).
Triterpenoid test was performed by giving 0.1 Table 1. The analysis result of B. asiatica seed extract based
gram of extract to 2 mL of ethanol 30%, heated on Thin Layer Chromatography and Column
and filtered. The filtrate was evaporated and ad- Chromatography
ded with eter by 1:1 ratio. The eter layer was ad- Obtained Thin Layer Column
ded with Lieberman Burchard reagent (three results Chromatography Chromatography
drops of acetic acid anhydride and one drop of Spot 3 2
concentrated H2SO4). The red colour revealed 0,24 0,60
the presence of triterpenoid. Rf Score 0,60 0,76
Analyses of Thin Layer Chromatography
and Column Chromatography The isolation and identification of triterpenoid
Thin Layer Chromatography was performed saponin from the methanol extract of B. asiatica
by following [13] method in which silica gel seeds with GCMS resulted in three Triterpenoid
adsorbent coated plate and methanol:chloroform: saponin compounds, namely 2.4-bis-(1.1-dime-
water (1:1:10) were treated as mobile phase. thylethyl)-, methylcarbamate; 4-Dodecylphenol;
Then, its Retension factor (Rf) was calculated by and 2.6 bis-(1.1-dimethylethyl)-4-methyl-, methyl-
dividing “the distance achieved by eluted compo- carbamate (Table 2; Picture 1).
unds” with “the distance achieved by elution li-
quid." The column chromatography was perfor- Table 2. The composition of triterpenoid compound in the
med by methanol: chloroform: water (1:1:10) sol- methanol extract of B. asiatica seeds with GC-MS
vent and silica gel column. The Rf analysis of co- Retention
N Abundance The possibility of
lumn chromatography was tested by thin layer time
o (%) compound name
1 14.300 2,61
ethyl)-, methylcarbamate
Analysis of GC-MS 2 18.850 1,13 4-Dodecylphenol
The extraction result of saponin was analyzed 2.6 bis-(1.1-
by GC-MS to determineits structure. The GC-MS 3 22.933 0,57 dimethylethyl)-4-methyl-
under following conditions: the injector tempera- , methylcarbamate

JTLS | J. Trop. Life. Science 120 Volume 4 | Number 2 | May | 2014

Meity et al, 2014

Picture 1. The GC MS result of methanol extract from B. asiatica seeds indicated three Triterpenoid saponin compounds

From these three compounds, the main com- 2.6 bis-(1.1-dimethylethyl)-4-methyl-, methylcar-
ponent was 2.4-bis-(1.1-dimethyl ethyl)-, bamate which are potential to be the bio-
methylcarbamate that had the biggest abundance pesticide.
of 2.61% with C14H22O mo-lecule for-mula.
Saponin consists of triter-pene glycoside and REFERENCES
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